Our broad, long term objective is to study tissue-specific regulation of salivary-type genes in human and/or macaque cell lines. The specific aim of this project is to determine conditions for primary cell culture of primate salivary gland cell lines as a prelude to establishing permanent cell lines that express salivary-type genes. We will employ a variety of culture methods to optimize conditions for cell survival, growth and differentiation in primary cultures of salivary gland epithelia. Macaque and human salivary gland tissues will be dispersed and established in a variety of culture media containing various hormones, growth factors and stimuli for secretion. Effects of calcium concentration, pH, CO2, O2, extracellular matrix components, feeder layers and growth in Methocel will also be tested. After establishing optimal conditions for primary culture of cells that express salivary-type genes, we will attempt to transform the cells in order to prepare useful permanent cell lines for further study of salivary-type gene regulation.